Cytokine Gene Expression in Splenic Macrophages and Kupffer Cells Following Haemorrhage

Abstract

TNF-alpha, IL-1, IL-6 and TGF-beta are important macrophage-derived mediators which play the pleiotropic role in inflammatory, metabolic, hematopoietic and immunologic processes. Studies have shown that haemorrhagic shock without significant tissue trauma induces profound immunosuppression which is associated with elevated plasma levels of TNF-alpha IL-1, IL-6 as well as TGF-beta. Furthermore, Kupffer cells but not the splenic M phi isolated from post-haemorrhaged animals showed an increased capacity to release inflammatory cytokines in response to LPS stimulation in vitro. However, it remains unknown whether the innate (i.e. in the absence of LPS stimulation) cytokine genes expression in Kupffer cells and splenic M phi is affected by haemorrhage. To determine this, C3H/HeN male mice were bled to and maintained at a mean arterial blood pressure of 35 mmHg for 60 min, and then adequately resuscitated. Splenic macrophages and Kupffer cells were isolated at 1 h after haemorrhage. Total RNA was extracted and cytokine mRNA was detected by semi-quantitative reverse transcription and polymerase chain reaction (RT-PCR). The results demonstrate that haemorrhage significantly elevated the mRNA accumulation of TNF-alpha, IL-1 beta, TGF-beta while IL-6 gene expression in Kupffer cells and splenic M phi was only slightly increased. Since Kupffer cells but not the splenic M phi showed increased cytokine release, it could be concluded that the differential regulation of cytokine release by these two macrophage populations following haemorrhage may be due to the divergence of the cytokine at the translational level.(ABSTRACT TRUNCATED AT 250 WORDS)